LOCUS       sequence-288718-        9125 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Insect Expression of DmNot4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9125)
  AUTHORS   Zekri L, Huntzinger E, Heimstadt S, Izaurralde E
  TITLE     The silencing domain of GW182 interacts with PABPC1 to promote 
            translational repression and degradation of microRNA targets and is 
            required for target release.
  JOURNAL   Mol Cell Biol. 2009 Dec;29(23):6220-31. doi: 10.1128/MCB.01081-09. 
            Epub 2009 Sep 21.
  PUBMED    19797087
REFERENCE   2  (bases 1 to 9125)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1128/MCB.01081-09"; journalName: "Mol Cell Biol"; date: 
            "2009-12"; volume: "29"; issue: "23"; pages: "6220-31"
COMMENT     Sequence Label: pAc5.1A-DmNot4-EGFP_F
FEATURES             Location/Qualifiers
     source          1..9125
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        56..2568
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     CDS             join(5805..5807,5808..5810,5811..6524)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(5805..5807,5808..5810,5811..6521)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5850..5871)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6111..6130)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6458..6479
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    6551..6685
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6575..6594)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6629..6648
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(7307..7895)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7387..7406)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8066..8857,8858..8926))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     8689..8708
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8927..9031)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     9099..9117
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cttcaaggaa ttaattctat attctaaaaa cacaaatgat acttctaaaa aaaatcatga
       61 atggcatcaa ctctgaatca aatctttgca gatgcaccta cttctcattt ccactgtcac
      121 atcatttttc cagatctcgc tgcctgttat gtggcccaca aaccaagaca cgttttatgg
      181 ccattaaagc tggctgatcg tcgccaaaca ccaaatacat atcaatatgt acattcgaga
      241 aagaagcgat caaagaagcg tcttcgggcg agtaggagaa tgcggaggag aaggagaacg
      301 agctgatcta gtatctctcc acaatccaat gccaactgac caactggcca tattcggagc
      361 aatttgaagc caatttccat cgcctggcga tcgctccatt cttggctata tgtttttcac
      421 cgttcccggg gccattttca aagactcgtc ggtaagataa gattgtgtca ctcgctgtct
      481 ctcttcattt gtcgaagaat gctgaggaat ttcgcgatga cgtcggcgag tattttgaag
      541 aatgagaata atttgtattt atacgaaaat cagttagtgg aattttctac aaaaacatgt
      601 tatctataga taattttgtt gcaaaatatg ttgactatga caaagattgt atgtatatac
      661 ctttaatgta ttctcatttt cttatgtatt tataatggca atgatgatac tgatgatatt
      721 ttaagatgat gccagaccac aggctgattt ctgcgtcttt tgccgaacgc agtgcatgtg
      781 cggttgttgt tttttggaat agtttcaatt ttcggactgt ccgctttgat ttcagtttct
      841 tggcttattc aaaaagcaaa gtaaagccaa aaaagcgaga tggcaatacc aaatgcggca
      901 aaacggtagt ggaaggaaag gggtgcgggg cagcggaagg aagggtgggg cggggcgtgg
      961 cggggtctgt ggctgggcgc gacgtcaccg acgttggagc cactcctttg accatgtgtg
     1021 cgtgtgtgta ttattcgtgt ctcgccactc gccggttgtt tttttctttt tatctcgctc
     1081 tctctagcgc catctcgtac gcatgctcaa cgcaccgcat gttgccgtgt cctttatgcg
     1141 tcattttggc tcgaaatagg caattattta aacaaagatt agtcaacgaa aacgctaaaa
     1201 taaataagtc tacaatatgg ttacttattg ccatgtgtgt gcagccaacg atagcaacaa
     1261 aagcaacaac acagtggctt tccctctttc actttttgtt tgcaagcgcg tgcgagcaag
     1321 acggcacgac cggcaaacgc aattacgctg acaaagagca gacgaagttt tggccgaaaa
     1381 acatcaaggc gcctgatacg aatgcatttg caataacaat tgcgatattt aatattgttt
     1441 atgaagctgt ttgacttcaa aacacacaaa aaaaaaaata aaacaaatta tttgaaagag
     1501 aattaggaat cggacagctt atcgttacgg gctaacagca caccgagacg aaatagctta
     1561 cctgacgtca cagcctctgg aagaactgcc gccaagcaga gagagagaga aaaagaggga
     1621 gagcagctta gaccgcatgt gcttgtgtgt gaggcgtctc tctcttcgtc tcctgtttgc
     1681 gcaaacgcat agactgcact gagaaaatcg attacctatt ttttatgaat gaatatttgc
     1741 actattacta ttcaaaacta ttaagatagc aatcacattc aatagccaaa tactatacca
     1801 cctgagcgat gcaacgaaat gatcaatttg agcaaaaatg ctgcatattt aggacggcat
     1861 cattatagaa atgcttcttg ctgtgtactt ttctctcgtc tggcagctgt ttcgccgtta
     1921 ttgttaaaac cggcttaagt taggtgtgtt ttctacgact agtgatgccc ctactagaag
     1981 atgtgtgttg cacaaatgtc cctgaataac caatttgaag tgcagatagc agtaaacgta
     2041 agctaatatg aatattattt aactgtaatg ttttaatatc gctggacatt actaataaac
     2101 ccactataaa cacatgtaca tatgtatgtt ttggcataca atgagtagtt ggggaaaaaa
     2161 tgtgtaaaag caccgtgacc atcacagcat aaagataacc agctgaagta tcgaatatga
     2221 gtaaccccca aattgaatca catgccgcaa ctgataggac ccatggaagt acactcttca
     2281 tggcgatata caagacacac acaagcacga acacccagtt gcggaggaaa ttctccgtaa
     2341 atgaaaaccc aatcggcgaa caattcatac ccatatatgg taaaagtttt gaacgcgact
     2401 tgagagcgga gagcattgcg gctgataagg ttttagcgct aagcgggctt tataaaacgg
     2461 gctgcgggac cagttttcat atcactaccg tttgagttct tgtgctgtgt ggatactcct
     2521 cccgacacaa agccgctcca tcagccagca gtcgtctaat ccagagaccc cggatcgggg
     2581 tacctactag tccagtgtgg tggaattctg cagatatcca gcacagtggc ggccgcgcca
     2641 acatgaacgg cctgagcagc aatgatgatg cagtcgagtg ccctttgtgt atggagccgc
     2701 ttgaggtgga cgatctgacc ttctttccgt gtacctgcgg atatcagatt tgccgattct
     2761 gttggcatag gatccgcacg gatgagaaca agctgtgtcc ggcatgccga aaggagtatc
     2821 ccgagaatcc agctgacttc aagccgctgt cgcaggagga aatgattgcc ttcaagtccc
     2881 agaaacgcca aagggaccaa caacgaaaac aaaagatcac cgagaaccgc aaacacttgg
     2941 ccaacgttcg cgttgtccaa aagaacttgg tgttcgttgt ggggctaccg ccacgacttg
     3001 ctgatgcaga catattaaag aaacacgagt atttcggtaa atatgggaaa attcataaag
     3061 tcgttataaa tccaagtacc acgtatgccg gggtccaggt aagagttggt ccatctgctt
     3121 ctgcgtatgt cacatatgtt aataactcgg atgccttgcg ggccattcaa agcgtcaata
     3181 atattatgat agacggtcgg cttataaaga ccagcttggg gacaaccaaa tactgtagcc
     3241 acttcatgaa gaaccagcag tgtcccaagg gcgactgcat gtacttacat gaattgggcg
     3301 atcccgaggc cagtttcaca aaggaggaaa tgcatcaggg aaagcacctg gagtacgaga
     3361 agcgcctgca cgatactcta attgcgtcat tgggaccgaa tgcaacagtc gcgattccat
     3421 cgtcatcgtc agcgtcatcc tcgtcctctg gatcgggtac aaatggttca agtgcatcag
     3481 gtaatgccca gcaaaaggag gcctggccaa gcttatcggt ctcgcccatc aatggcaaag
     3541 aggcggctgc cactgcgacc agttccagtg ggaaaagtaa acgggaaaag ctgcgcaacg
     3601 agaagaggca cgagaagaac aaggcgaaga ataagaacgg cagcaataca aacgccaatg
     3661 cctcgaacaa ggagaactat gttcccgaaa cgagaagcag cacaagtact gagacattcg
     3721 cagaggctac ggcggatgca ccggcttcca ccaaggcaga accgccgcaa gcctctagca
     3781 atagaacgag ggcggatcgt ggaaaagatc ggaccacggc tagtgcaaag gagcagaaga
     3841 agagcaagga agctgctcca gcacctgcag caagtaaacc ggcggagcgg gttgaaacaa
     3901 gcgagagtac aataagacaa aagaaggcgg aagtaacaga aagctgtgaa gataacttac
     3961 cacaaaagag attagcggga acaaacgttc aaagatctgt gagctcttgt agcgaaaata
     4021 gcgaaggaca cgtctctgag agtagcttaa gtgagaagag tttaactggt gattatgtgg
     4081 aggaaaagtg caatagtgtg aattcggaaa gccagcaaga aagtgtaaag tttcaggagg
     4141 agctcgaaaa gagcaacgag gcaattgtcg aggccgaaac gattctgcca acagctgagt
     4201 cctctgagga catcagtccc gctgcagttg cgccaggcaa tggagaagtc gaaggatgct
     4261 tacccgttgt tgatccagtc gaaccaccga gtctggcgga taatggaagc agagtaacag
     4321 atgcgctatc taagctgaac attttcgatg acacgcctag ctttttcaca tcgccatcat
     4381 tccagcaagc acccatatta aagaataagc tagatttgga aatgcgacag tctcatttac
     4441 ctgacttggt caacgacatt gatggaatcc aaaaggcatc caatacaaac gagtgggagg
     4501 aagccttcaa aaatgtgatg atgggcaaca cgcagcacat ggaggagcaa ctgttgcagc
     4561 agcaacattt gcagcagcat cagaatttgc gccaccagct ggtgttgcaa caggaggagt
     4621 tccttcgcat gcaagaatta cagaaacgca acaactttgc gacgcaaatc aacggtcctg
     4681 cgaatgattt ccttagagct tatgagctca gagctcaagc caatgcaatc attcagcaac
     4741 aattattaca acagcacgca ggcgaaaatc tatttggcgg caacatgtcg aagttcttcg
     4801 atttccataa aagtcagccg cagtcacacc atcagtatct aaatggacat ccgcctcaaa
     4861 tcaatgggaa cggtgctgtg ccggagccgc aaagagtggc ggcctctttg gaaagcaatc
     4921 gactgaattc gccttttgtc gaaaatggac ttattaactc gcagcagcag cagcagcagc
     4981 agcaacagca acaaaagcag agaatgatgg gaatgtatga atttatgcct ccaaacacgc
     5041 aatctcagca gaatcggttt acacagaact cgatagtcga cgatgactta ggattcgacc
     5101 cgttcgttga gacccaaaag ggacttgctg aacttatgga aaatgaggtg gtccaaaaac
     5161 agagtattaa taatgagaac cccttgccga agttgccgcc acagcctcaa gttccacccc
     5221 atccgcaatt ggtggacaac ctgcagcgag ctcgcatgcc gccgccaggc ttcaatcacg
     5281 tcaacacatt gggcttgggc ggtgcatcca gactgcagct caccagcaaa ataataccct
     5341 tcatgaacat gcccgtcaat ggggtgggca acagcggtgc ccagggacag caccaaatgc
     5401 caatgggtgt caactggaat gcgccgatgg gcatgcacca gaacccggga cagcccgtgg
     5461 gtgattcgca attgcagcat ccaatggctc acaacaaggt ttacaacaac agtgattgga
     5521 cttcaatgga tccggctata ctttcgttta gacagtattc ttcttttcca caaaaccaaa
     5581 ttcccccaca tcctcagcag caacaggatt tatttttgca gcatttggct caacagcaaa
     5641 attcccagag tggtggtttc aacaaccagg cacagcaaat gcttcctatg gggatgccca
     5701 atagtttgct gaacggacaa caaacgcagc cgccacaggt caatgccaat gttcagggca
     5761 tgcttgagtt tttaaaaagc cgtcaattcg ttttcgaagc caacatggtg agcaagggcg
     5821 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     5881 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     5941 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga
     6001 cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca
     6061 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     6121 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     6181 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     6241 acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact
     6301 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
     6361 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
     6421 ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     6481 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaagagctc gctgatcagc
     6541 ctcgatcgag gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag
     6601 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     6661 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     6721 tcagggggag gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtatggc
     6781 tgattatgat cagtcgaccg atgcccttga gagccttcaa cccagtcagc tccttccggt
     6841 gggcgcgggg catgactatc gtcgccgcac ttatgactgt cttctttatc atgcaactcg
     6901 taggacaggt gccggcagcg ctctgggtca ttttcggcga ggaccgcttt cgctggagcg
     6961 cgacgatgat cggcctgtcg cttgcggtat tcggaatctt gcacgccctc gctcaagcct
     7021 tcgtcactgg tcccgccacc aaacgtttcg gcgagaagca ggccattatc gccggcatgg
     7081 cggccgacgc gctgggctac gtcttgctgg cgttcgcgac gcgaggctgg atggccttcc
     7141 ccattatgat tcttctcgct tccggcggca tcgggatgcc cgcgttgcag gccatgctgt
     7201 ccaggcaggt agatgacgac catcagggac agcttcaagg atcgctcgcg gctcttacca
     7261 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     7321 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     7381 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     7441 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     7501 aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     7561 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     7621 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     7681 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     7741 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     7801 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     7861 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     7921 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     7981 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     8041 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     8101 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     8161 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     8221 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     8281 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     8341 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tgcaggcatc gtggtgtcac
     8401 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     8461 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     8521 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     8581 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     8641 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aacacgggat aataccgcgc
     8701 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     8761 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     8821 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     8881 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     8941 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     9001 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     9061 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     9121 ttcgt
//
